Expression of G1- epitope of bovine ephemeral fever virus in E. coli : A novel candidate to develop ELISA kit
نویسندگان
چکیده
Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G of bovine ephemeral fever virus (BEFV) is composed of 4 antigenic sites (G1-G4) and plays the main role for eliciting neutralizing antibodies and protective immunity. The G1 - epitope is a linear antigenic site and conserved among BEFV strains. In order to develop an ELISA test based on G1-epitope as coating antigen, this study was carried out to express the recombinant G1-epitope of BEFV in prokaryotic system. Using PCR and specific primers, a length of 88 amino acid of the G glycoprotein of BEFV including G1- epitope was amplified and cloned into the expression vector pGEX-4T-1, with the GST moiety. The recombinant plasmid (pGEX-4T-1-G1) was then transformed into Escherichia coli BL21 and expression of fusion protein was induced by 0.10 mM IPTG. The maximum expression of the fusion protein was obtained at 16 hr post induction as verified by SDS-PAGE electrophoresis, and it was also confirmed that this protein bearing G1- epitope is sufficiently biologically active to bind to anti-BEFV serum in western blot experiment.
منابع مشابه
Expression of G1- epitope of bovine ephemeral fever virus in E. coli : A novel candidate to develop ELISA kit
Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G of bovine ephemeral fever virus (BEFV) is composed of 4 antigenic sites (G1-G4) and plays the m...
متن کاملProduction of Monoclonal Antibody against Prokaryotically Expressed G1 Protein of Bovine Ephemeral Fever Virus
Epitope-G1 of bovine ephemeral fever virus (BEFV) G glycoprotein has been genetically and antigenically conserved among various isolates of BEFV and only reacts with anti-BEFV neutralising antibodies. Therefore, it is a candidate antigen for development of the enzyme linked immunosorbent assay (ELISA) for serological identification bovine ephemeral fever (BEF)-infected animals. The aim of this ...
متن کاملCloning and Expression of the G1 Epitope of Bovine Ephemeral Fever Virus G Glycoprotein in Escherichia Coli
Bovine ephemeral fever (BEF) is an acute epidemic disease in cattle and water buffalo, spanning tropical and subtropical zones of Asia, Australia, and Africa. In recent years, BEF has been distributed in many provinces of Iran and caused economic losses. In this study the sequence encoding the G1 epitope of bovine ephemeral fever virus (BEFV) G glycoprotein was amplified by PCR, ligated into th...
متن کاملCloning, Expression and Purification of a Novel Multi-epitopic HIV-1 Vaccine Candidate: A Preliminary Study on Immunoreactivity
Introduction : Designing an effective vaccine against human immunodeficiency virus (HIV)-1 is a global health priority . Multi-epitope vaccines offer several potential advantages that may be promising in case of mutable divergent pathogens such as HIV-1. Herein, a multiepitopic recombinant protein containing various HIV-1 antigens was expressed in E. coli cells and its immunogenicity in combi...
متن کامل